USPTO Patents Application 09905777

U.S. S.N 09/905,777
Chen

PRELIMINARY AMENDMENT

enabling safe transcutaneous photoactivation. While there are several reports in

the scientific literature of utilizing the specificity of the binding between biotin

and streptavidin to target tumor cells, there are no reports utilizing this ligand-

receptor binding pair aimed at vascular lesions nor in conjunction with prolonged

PDT light exposure (see, for example: Savitsky eta/., SP/E, 3191: 343-

353,1997; and Ruebner et a/., SP/E, 2625: 328-332, 1996). In a non-PDT

modality, the biotin-streptavidin-receptor binding pair has also been used as

tumor targeting conjugates with radionuclides (see: U.S. Pat. No. 5,630,996

(Reno eta/.) and with monoclonal antibodies (see: Casalini et a/; J. Nuclear

Mec/., 38(9): 1 378-1 381 ,J 997) and U.S. Pat. No. 5,4&2,698 (Griffiths)).

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Please replace the paragraph on page 7, line 20, through page 8, line 3,

with the following paragraph:

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A still further embodiment of this invention is drawn to a method for
transcutaneous photodynamic therapy of target lesion in a mammalian subject
comprising: administering to the subject a therapeutically effective amount of a
first conjugate comprising a first member of a ligand-receptor binding pair
conjugated to an antibody or antibody fragment, where the antibody or antibody
fragment selectively binds to a target antigen found on thick or thin neointimas,
arterial plaques, vascular smooth muscle cells and/or the abnormal extracellular
matrix of the site to be treated. This step is followed by administering to the
subject a therapeutically effective amount of a second conjugate comprising a
second member of the ligand-receptor binding pair conjugated to a
photosensitizing agent or photosensitizing agent delivery system or prodrug,
where the first member binds to the second member of the ligand-receptor
binding pair. A subsequent step includes irradiating at least a portion of the
subject with light at a wavelength or waveband absorbed by the
photosensitizing agent or if prodrug, by the product thereof. This embodiment
further includes that the light is provided by a light source and that the
irradiation is at a relatively low fluence rate that results in the activation of the
photosensitizing agent or prodrug product.

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U.S. S.N 09/905,777
Chen

PRELIMINARY AMENDMENT

Please replace the paragraph on page 8, lines 16-28, with the following

paragraph:

Yet another embodiment of the present invention is drawn to a method
for transcutaneous ultrasonic therapy of a target lesion in a mammalian subject
comprising:

administering to the subject a therapeutically effective amount of an
ultrasonic sensitizing agent or a ultrasonic sensitizing agent delivery system or a
prodrug, where the ultrasonic sensitizing agent or ultrasonic sensitizing agent
delivery system or prodrug selectively binds to the thick or thin neointimas,
arterial plaques, vascular smooth muscle cells and/or the abnormal extracellular
matrix of the site to be treated. This step is followed by irradiating at least a
portion of the subject with ultrasonic energy at a frequency that activates the
ultrasonic sensitizing agent or if a prodrug, by a prodrug product thereof, where
the ultrasonic energy is provided by an ultrasonic energy emitting source. This
embodiment further provides that the ultrasonic therapy drug is cleared from

non-target tissues of the subject prior to irradi^fon.

Please replace the paragraph on page 4. lines 13-19, with the following -
paragraph:

Other embodiments of the present invention areTdra wn ToTl^prSggrrny^^
disclosed methods of transcutaneous PDT, where the light source is positioned
in proximity to the target tissue of the subject and is selected from the group
consisting of: an LED light source; an electroluminescent light source; an
incandescent light source; a cold cathode fluorescent light source; organic
polymer light source; and inorganic light source. A preferred embodiment

= =JiicdudesJ^ Nght source.

Please replace the paragraph on page 11, line 25, through page 12, line
1, wit h the following para graph:

Further, as used herein "target cells" or "target tissues" are those cells or
tissues, respectively that are intended to be impaired or destroyed by this
treatment method. Target cells or target tissues take up the photosensitizing

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U. S.S.N 09/905,777
Chen

PRELIMINARY AMENDMENT

agent; then when sufficient radiation is applied, these cells or tissues are
impaired or destroyed. Target cells are those cells in target tissues, which
include, but are not limited to: vascular lesions, thick or thin neointimas, arterial
plaques, neoplasms, vascular smooth muscle cells and the abnormal
extracellular matrix of the site to be treated. "Non-target cells" are all the cells
of an intact animal which are not intended to be impaired or destroyed by the
treatment method. These non-target cells include but are not limited to healthy

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nf 1 Qth^-D-QJmal ti ssue, not otherwis e identified to J >e^targeted .

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Please replace the paragraph on page 1 2, lines 23-28, with the following
paragraph:

"Radiation" as used herein includes all wavelengths. Preferably, the
radiation wavelength is selected to match the wavelength(s) which excites the
photosensitive compound. Even more preferably, the radiation wavelength
matches the excitation wavelength of the photosensitive compound and has low
absorption by the non-target cells and the rest of the intact animal, including
blood proteins. For example, the preferred wavelength for ICG is the range of
750-850 nm.

Please replace the paragraph on page 13, line 29, through page 14, line
3 r w'*h thfl following paragraph:

The photosensitizing agent also can be conjugated to specific ligands
reactive with a target, such as receptor-specific ligands or immunoglobulins or
immunospecific portions of immunoglogulins, permitting them to be more
concentrated in a desired target cell or microorganism. The photosensitizing
agent may be further conjugated to a ligand-receptor binding pair, which
includes, but is not limited to: biotin-streptavidin; and antigen-antibody. This
conjugation may permit lowering of the required dose level since the material is
more selectively targeted and less is wasted in distribution into other tissues
whose destruction must be avoided.

j Please replace the paragraph on page 14fline 20, through page TBT^lin
5, with the following paragraph:

4

U. S.S.N 09/905,777
Chen

PRELIMINARY AMENDMENT

This method comprises irradiating at least a portion of the subject with
light at wavelength or waveband absorbed by said photosensitizing agent that
under conditions of activation during photodynamic therapy using a relatively
low fluence rate, but also at an overall high total fluence dose resulting in
minimal collateral tissue damage. It is contemplated that the optimal total
fluence will be determined clinically using a light dose escalation trial. It is
further contemplated that the total fluence will preferably be in the range of 30
Joules to 25,000 Joules, and more preferably be in the range from 100 Joules
to 20,000 Joules, and most preferably be in the range from 500 Joules to
10,000 Joules. The methods comprise irradiating at least a portion of the
subject with light at a wavelength or waveband absorbed by said
photosensitizing agent that under conditions of activation during photodynamic
therapy using a relatively low fluence rate, but an overall high total fluence dose
resulting in minimal collateral normal tissue damage. What is meant by
"relatively low fluence rate" is a fluence rate that is lower than that typically
used and one that generally does not result in significant damage to collateral or
non-target tissues. Specifically, the intensity of radiation used to treat the
target cell or target tissue is preferably between about 5 and 100 mW/cm 2 .
More preferably, the intensity of radiation is between about 10 and 75 mW/cm 2 .
Most preferably, the intensity of radiation is between about 15 and 50 mW/cm 2

Please replace the paragraph on page 16fHne 2fffthrough page^T7fHne
T2, with the following par agraph :

The ordinary skilled artisan would be familiar with various ligand-receptor
binding pairs, including those known and those currently yet to be discovered.
Those known, include, but are not limited to the group consisting of: biotin-
streptavidin; and antigen-antibody. This invention contemplates a preferred
embodiment that includes the use of biotin-streptavidin as the ligand-receptor
binding pair. However, the ordinary skilled artisan would readily understand
from the present disclosure that any ligand-receptor binding pair may be useful
provided the ligand-receptor binding pair demonstrate a specificity for the

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U.S. S.N 09/905,777
Chen

PRELIMINARY AMENDMENT

binding by the ligand to the receptor and further provided that the ligand-
receptor binding pair permit the creation of a first conjugate comprising a first
member of the ligand-receptor binding pair conjugated to an antibody or
antibody fragment, wherein said antibody or antibody fragment selectively binds
to a target antigen of thick or thin neointimas, arterial plaques, vascular smooth
muscle cells and/or the abnormal extracellular matrix of the site to be treated;
and further permit the creation of a second conjugate comprising a second
member of the ligand-receptor binding pair conjugated to an energy sensitizing
or photosensitizing agent or energy sensitizing or photosensitizing agent delivery
system or prodrug, and further wherein the first member binds to the second
member of the ligand-rec eptor binding pair.

Please replace the paragraphs on page 19, line 24, through page 20, line
4, with the following paragraphs:

The PDT light source is an externally positioned light source
directed at the site to be treated. The light source may be a laser diode (2),
light emitting diode or other electroluminescent device. The light source is
angled and the light beam is focused so as to direct the light through the skin
(3) or membrane of the mammalian subject being treated in a direction
lengthwise and parallel to the vessel wall (5) to plaque (4). See Figures 1A and
1B.

Alternatively, the light source could comprise a laser diode (2) coupled to
an optical fiber (6) which is then aimed at the vessel so as to direct the light
along the length of the vessel. See Figure 2. The light source could also
comprise a strip of light emitting diodes (LEDs) (7) which are then arrayed on
the skin or the membrane overlying the site to be treated in the mammalian
subject. See Figure 3. The light source could also comprise an optical fiber
diffuser (8) which is placed over the skin or the membrane overlying the site to
be treated in the mammalian subject. See Figure 4. A mirrored surface (9) may
direct light downward.